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Figure 1: Western blot analysis of U251-MG and PC3 cell line extracts using Tumor Protein P53 (TP53) antibody (Catalog number ABIN967416, Lot number 2335876). TP53 is present in the positive control sample (U251-MG) and absent from the negative control sample (PC3). The arrowhead indicates the expected position of TP53 (predicted MW ~53kDa). 32 micrograms of total protein lysates from each sample were loaded into each lane. Upper panel: scanned image of the TP53 antibody probed with the U251-MG and PC3 extracts in lanes 1 and 2 respectively. Lower panel: scanned image of the loading control (beta-actin).

P53 is a 53 kD nuclear phosphoprotein that acts as a tumor suppressor protein, and is involved in inhibiting cell proliferation when DNA damage occurs. The gene for p53 is the most commonly mutated gene yet identified in human cancers. Missense mutations occur in tumors of the colon, lung, breast, ovary, bladder and several other organs. The mutant p53 is overexpressed in a variety of transformed cells and the wildtype p53 forms specific complexes with several viral oncogenes including SV40 large T, E1B from adenovirus and E6 from human papilloma virus. Wildtype p53 plays a role as a checkpoint protein for DNA damage during the S-phase of the cell cycle. p53 migrates at a reduced molecular weight of 53 kDa. Clone G59-12 recognizes mutant and wild type human, rat and mouse p53 tumor suppressor protein. Recombinant full-length human p53 was used as immunogen.

13. The membrane was rinsed three times with PBS containing 0.05% Tween-20.4. Incubated in Acidic Glycine Stripping Buffer at room temperature with gentle agitation for 3 times, 10

min each.

15. The membrane was washed in PBS containing 0.05% Tween-20 times for 10 min each.

16. Repeated Steps 5-12 with the loading control antibody (beta-actin) and its matching secondary

antibody.

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Figure 1: Western blot analysis of U251-MG and PC3 cell line extracts using Tumor Pro...

Figure 1: Western blot analysis of U251-MG and PC3 cell line extracts using Tumor Protein P53 (TP53) antibody (Catalog number ABIN967416, Lot number 2335876). TP53 is present in the positive control sample (U251-MG) and absent from the negative control sample (PC3). The arrowhead indicates the expected position of TP53 (predicted MW ~53kDa). 32 micrograms of total protein lysates from each sample were loaded into each lane. Upper panel: scanned image of the TP53 antibody probed with the U251-MG and PC3 extracts in lanes 1 and 2 respectively. Lower panel: scanned image of the loading control (beta-actin).

Figure 1: Western blot analysis of U251-MG and PC3 cell line extracts using Tumor Protein P53 (TP53) antibody (Catalog number ABIN967416, Lot number 2335876). TP53 is present in the positive control sample (U251-MG) and absent from the negative control sample (PC3). The arrowhead indicates the expected position of TP53 (predicted MW ~53kDa). 32 micrograms of total protein lysates from each sample were loaded into each lane. Upper panel: scanned image of the TP53 antibody probed with the U251-MG and PC3 extracts in lanes 1 and 2 respectively. Lower panel: scanned image of the loading control (beta-actin).